Abstract
The GFP-based superecliptic pHluorin (SEP) enables detection of exocytosis and endocytosis, but its performance has not been duplicated in red fluorescent protein scaffolds. Here we describe "semisynthetic" pH-sensitive protein conjugates with organic fluorophores, carbofluorescein, and Virginia Orange that match the properties of SEP. Conjugation to genetically encoded self-labeling tags or antibodies allows visualization of both exocytosis and endocytosis, constituting new bright sensors for these key steps of synaptic transmission.
Publication types
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Research Support, N.I.H., Intramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Biosensing Techniques / methods
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Drug Design
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Endocytosis*
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Exocytosis*
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Fluoresceins / chemical synthesis
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Fluoresceins / chemistry
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Fluorescent Dyes* / chemical synthesis
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Fluorescent Dyes* / chemistry
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Green Fluorescent Proteins / chemistry
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Hippocampus / cytology
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Hippocampus / metabolism
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Hydrogen-Ion Concentration
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Luminescent Proteins / chemistry
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Neurons / metabolism
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PC12 Cells
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Rats
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Red Fluorescent Protein
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Synaptic Transmission
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Synaptic Vesicles / physiology
Substances
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Fluoresceins
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Fluorescent Dyes
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Luminescent Proteins
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PHluorin
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Green Fluorescent Proteins